ATP-synthesis by proteoliposomes incorporating Rhodospirillum rubrum F0F1 as measured with firefly luciferase: dependence on delta psi and delta pH.

ATP-synthesis catalyzed by proteoliposomes incorporating Rhodospirillum rubrum F0F1 was driven by artificially applied electrochemical proton gradients. The time-course of ATP-synthesis was followed continuously by means of firefly luciferase. Correction methods were developed which allow one to calculate the initial rate of ATP-synthesis from the observed luminescence kinetics. The following results were obtained: (1) ATP-synthesis occurred above a threshold delta mu H+ of 90 mV; this threshold is not imposed by the activation requirement of the enzyme; (2) delta psi and delta pH appear to be equivalent as driving forces for ATP-synthesis if allowance is made for the effect of the electrical capacitance of the liposome membrane on the distribution of K+ at equilibrium; and (3) the highest rate observed so far is 200 mol ATP per mol F0F1 per s.